Technical Data
A1309-52
Albumin, BCP (Bromcresol Purple) Assay Kit, BioAssay™
1Kit
Kits and Assays Storage: 4°C/-20°CShipping: RT
Albumin is the most abundant plasma protein in human. It accounts for about 60% of the total serum protein. Albumin plays important physiological roles, including maintenance of colloid osmotic pressure, binding of key substances such as long-chain fatty acids, bile acids, bilirubin, haematin, calcium, magnesium. It has anti-oxidant and anticoagulant effects, and also acts as a carrier for nutritional factors and drugs, as an effective plasma pH buffer. Serum albumin is a reliable prognostic indicator for morbidity and mortality, liver disease, nephritic syndrome, malnutrition and protein-losing enteropathies. High levels are associated with dehydration.

Simple, direct and automation-ready procedures for measuring albumin concentration in biological samples are becoming popular in Research and Drug Discovery. BCP albumin assay kit is designed to measure albumin directly in biological samples without any pretreatment. The improved method utilizes bromcresol purple that forms a colored complex specifically with albumin. The intensity of the color, measured at 610nm, is directly proportional to the albumin concentration in the sample. The optimized formulation substantially reduces interference by substances in the raw samples.

Key Features:
Sensitive and accurate. Use as little as 20ul samples. Detection range 0.3 g/dL (45uM) to 5 g/dL (750uM) albumin in 96-well plate assay.
Simple and high-throughput. The procedure involves addition of a single working reagent and incubation for 5 min. Can be readily automated as a high-throughput assay for thousands of samples per day.
Improved reagent stability and versatility. The optimized formulation has greatly enhanced reagent and signal stability. Cuvet or 96-well plate assay.
Low interference in biological samples. No pretreatments are needed. Assays can be directly performed on raw biological samples i.e., in the presence of lipid and protein.

Applications:
Direct Assays: albumin in serum, urine, biological preparations.
Drug Discovery/Pharmacology: effects of drugs on albumin metabolism.

Kit Contents:
1. Reagent, 1x50ml
2. Albumin standard (5g/dL), 1x2ml

Storage and Stability:
The kit is shipped at room temperature. Store Reagent at 4°C and Albumin standard at -20°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. 

Procedures:
Reagent Preparation:
Important: bring reagents to room temperature and shake before use.

Procedure using 96-well plate:
1. Dilute standards in distilled water as follows. Dilute serum samples 2 fold. Transfer 20ul diluted standards and diluted samples to wells of a clear bottom plate. Store diluted standards at -20°C for future use.
No STD+H2O Vol (ul) BSA (g/dL)
1 100ul+0ul 100 5.0
2 80ul+20ul 100 4.0
3 60ul+40ul 100 3.0
4 40ul+60ul 100 2.0
5 30ul+70ul 100 1.5
6 20ul+80ul 100 1.0
7 10ul+90ul 100 0.5
8 0ul+100ul 100 0
2. Add 200ul working reagent and tap lightly to mix. Avoid bubble formation!
3. Incubate 5 min at room temperature and read optical density at 590- 630nm (peak absorbance at 610nm).
Procedure using cuvette:
1. Transfer 60ul Blank, Standards and samples to appropriately labeled tubes. Add 1000ul working reagent and tap lightly to mix. Incubate 5 min at room temperature.
2. Transfer to cuvet and read optical density at 610nm.
Important: if sample OD is higher than the OD for standard, dilute samples in distilled water and repeat the assay.

Calculation:
Subtract blank OD (water, #8) from the standard OD values and plot the OD against standard concentrations. Use the standard curve to determine the sample albumin concentration.
Conversions: 0.1 g/dL albumin equals 15uM, 0.1% or 1000 ppm.

Materials Required, But Not Provided:
Pipeting devices and accessories (e.g. 5ul).
Procedure using 96-well plate:
Clear bottom 96-well plates (e.g. Corning Costar) and plate reader.
Procedure using cuvette:
Spectrophotometer and cuvets for measuring OD at 610nm.

Examples:
Albumin was assayed in duplicate using the 96-well assay protocol. The albumin content (g/dL) was 3.6 ± 0.0, 2.8 ± 0.1 and 4.3 ± 0.0 in rat serum, fetal bovine serum and goat serum, respectively. The albumin content in a fresh human urine sample was below the detection limit (0.3 g/dL).
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
1. Maier SM et al (2007) Proteinuria of Nonautoimmune Origin in Wild-type FVB/NJ Mice. Comp Med. 57(3):255-66.
2. Sharifuzzaman SM and Austin B (2009). Kocuria SM1 controls vibriosis in rainbow trout (Oncorhynchus mykiss, Walbaum). J Appl Microbiol. 108(6): 2162 – 2170.
3. Shin SY et al (2009). Immunological investigation in the adenoid tissues from children with chronic rhinosinusitis. Otolaryngol Head Neck Surg. 141(1):91-6.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.